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Updated: Aug 15, 2026

Uracil-DNA Glycosylase Assay by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis
Published on: April 22, 2022
Mutation detection in DNA oligonucleotides based on a Guanine quenching method coupled with enzymatic digestion of
Tatsuo Maruyama1, Toshimitsu Shinohara, Hirofumi Ichinose
1Department of Applied Chemistry Graduate School of Engineering, Kyushu University, 6-10-1 Hakozaki, 812-8581, Fukuoka, Japan.
Abstract:
Fluorescence quenching by guanine allows DNA hybridization to be monitored and any point mutations in oligonucleotides to be detected. However, fluorescence quenching is often affected by untargeted guanine located in a protruding end (single-strand DNA) of the probe-target DNA duplex resulting in an unsatisfactory sensitivity. In the present study, we used enzymatic digestion of the protruding end of a probe-target DNA duplex to avoid interference by untargeted guanine on fluorescence quenching for detection of a nucleobase mutation. Enzymatic digestion of the protruding end of the DNA duplex fully prevented interference by untargeted guanine, and produced a marked difference in the quenching ratios (36% for wild-type, and 0% for mutant).
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